Antibody and protein formulations

ABSTRACT

Provided are salt-free antibody and other protein formulations that are substantially isosmotic and of low viscosity. Also provided are methods for the treatment of diseases using the disclosed formulations.

BACKGROUND

The present disclosure relates generally to antibody and other proteinformulations that are isosmotic and of low viscosity, includingformulations that are useful for injection and general administration.

Hemophilia patients have bleeding disorders that result in delayed bloodclotting after an injury or surgery. Prolonged bleeding is caused by agenetic deficiency in one or more blood clotting factor. Two commontypes of hemophilia are known—hemophilia A and hemophilia B. HemophiliaA is caused by a deficiency in factor VIII whereas hemophilia B iscaused by a deficiency in factor IX. About 75-80% of total hemophiliapatients have hemophilia A.

Tissue factor pathway inhibitor (TFPI) is a human inhibitor of theextrinsic pathway of blood coagulation and functions in anticoagulation.Antibodies that are directed against TFPI, including anti-TFPImonoclonal antibodies (aTFPI mAb), are being developed in an effort toblock TFPI function. One such aTFPI mAb is a human IgG2 anti-TFPI mAb.that is being developed for the treatment of hemophilia A and Bpatients.

Antibody and other proteins may be administrated to patients viaintravenous, intramuscular, and/or subcutaneous injection. To ensurepatient compliance, it is desirable that subcutaneous injection dosageforms be isotonic and include small injection volumes (<2.0 ml perinjection site). To reduce injection volume, proteins are oftenadministered within the range of 1 mg/ml to 150 mg/ml.

While both liquid and lyophilized dosage forms are used for currentlymarketed antibody and other protein-based drug products, lyophilizedforms are more frequently used for protein and antibody drug productshaving high protein concentrations.

A protein and antibody dosage form may present many challenges informulation development, especially for liquid formulation. Forformulations in which the protein concentration is near its apparentsolubility limit, phase separation can occur through precipitation,gelation, and/or crystallization. At high protein concentration, thestability of an antibody or other protein can become problematic due tothe formation of soluble and insoluble protein-protein aggregates.Highly concentrated protein formulations are frequently highly viscous,which presents difficulties for processing, such as ultrafiltration andsterile filtration, and for injection of the dosage solution. And atprotein concentrations that are desirable for formulations intended forintramuscular or subcutaneous administration, high concentrations ofstabilizers, such as sucrose and sodium chloride, are required toachieve long-term protein stability. The resulting hypertonic solutionsoften cause injection pain due to tissue damage. Therefore, it iscritical to balance the amount of stabilizers for stability andosmolality of the high protein concentration formulation.

For these reasons, there is a need in the art for antibody and otherprotein-based therapeutic formulations in liquid form that exhibit highprotein concentrations without the problem of significantly increasedprotein aggregation, osmolality, or viscosity and/or decreased proteinstability. It is, therefore, desirable that antibody and otherprotein-based formulations contain limited amounts of excipients andsmall volumes for ease of therapeutic administration or delivery. It isfurther desirable that antibody and other protein-based therapeuticformulations be amenable to lyophilization to enhance protein stabilityunder prolonged storage conditions.

SUMMARY

The present disclosure provides liquid and lyophilized antibody andprotein-based formulations that are substantially isotonic and lowviscosity and that contain substantially no inorganic salt. The antibodyand other protein formulations presented herein contain from about 0 mMto about 30 mM histidine; from about 50 ppm to about 200 ppm of anon-ionic surfactant such as, for example, polysorbate (Tween®) 80 orpolysorbate (Tween®) 20; from about 88 mM to about 292 mM of a sugar orsugar alcohol such as, for example, mannitol, dextrose, glucose,trehalose, and/or sucrose; from about 0 mM to about 50 mM arginine; fromabout 0 mM to about 50 mM lysine; from about 0 mM to about 133 mMglycine or alanine; from about 0 mM to about 10 mM methionine; and fromabout 1 mg/ml to about 150 mg/ml of a protein at a pH from about pH 4.0to about pH 6.0. The formulations disclosed herein exhibit a viscosityranging from about 1 mPa-S to about 20 mPa-S at 22° C., or from about 1mPa-S to about 15 mPa-S at 22° C., or from about 1 mPa-S to about 10mPa-S at 22° C. or from about 1 mPa-S to about 8 mPa-S at 22° C. or fromabout 1 mPa-S to about 6 mPa-S at 22° C. and osmolality ranging fromabout 240 to about 380 mmol/kg.

Within further aspects, the present disclosure provides methods for thetreatment of a disorder in a patient, comprising the administration tothe patient of a therapeutically effective amount of one or moreformulations described herein. For example, provided are methods for thetreatment of a disorder in a patient, comprising the administration tothe patient of a therapeutically effective amount of an antibody orother protein formulation as described in greater detail herein.

These and other features of the present teachings are set forth herein.

BRIEF DESCRIPTION OF THE DRAWINGS

The skilled artisan will understand that the drawings, described below,are for illustration purposes only. The drawings are not intended tolimit the scope of the present teachings in any way.

FIG. 1 is a graph showing the effect of sodium chloride (NaCl)concentration on the turbidity of 20 mg/ml anti-TFPI mAb formulations atpH 5.5.

FIG. 2 is a graph showing the effect of pH on the turbidity of ananti-TFPI mAb drug substance.

DESCRIPTION OF VARIOUS EMBODIMENTS

As described above, the present disclosure provides antibody and otherprotein formulations that stabilize the antibody or other protein inliquid form or lyophilized form at intended storage conditions. Theformulations described herein include one or more pharmaceuticallyacceptable excipients or stabilizers, and are contained in bufferedmedia at a suitable pH and are substantially isosmotic withphysiological fluids. For systemic administration, injection is oneroute of administration, including intramuscular, intravenous,intraperitoneal, and subcutaneous injection.

Because of their low viscosity, the presently disclosed proteinformulations can be conveniently processed via, for example,ultrafiltration and sterile filtration and can be administered to apatient via injection, including subcutaneous injection. Moreover,because they are substantially isosmotic, the presently disclosedantibody and protein formulations reduce tissue damage or other adversephysiologic effects and thereby achieving favorable patient toleranceand increased patient compliance.

The formulations described herein are characterized by the substantialabsence of added salt, which provides the flexibility for increasing theconcentrations of other stabilizers, such as sucrose, while maintainingthe osmolality of the formulation for improved in vivo tolerability and,consequently, increased patient compliance. Moreover, the low viscosityof the presently described formulations permits convenient processing,including but not limited to ultrafiltration and sterile filtration, andinjection of the drug product solution through the needle.

As used herein, the term “viscosity” refers to the resistance of aliquid formulation to flow, such as when injected through a syringeneedle during administration to a patient. Viscosity measurements can bedone by a cone and plate technique with a Peltier element set at adefined temperature, such as 22° C. as described herein. Typically, awell-defined shear stress gradient is applied to the liquid formulationand the resulting shear rate is measured. The viscosity is the ratio ofthe shear stress to the shear rate. As used herein, viscosity isexpressed in units of mPa-S at 22° C. wherein 1 mPa-S=1 cP. The lowviscosity, substantially isosmotic formulations disclosed herein aretypically characterized by having a viscosity ranging from about 1 mPa-Sto about 20 mPa-S at 22° C., or from about 1 mPa-S to about 15 mPa-S at22° C., or from about 1 mPa-S to about 10 mPa-S at 22° C. or from about1 mPa-S to about 8 mPa-S at 22° C. or from about 1 mPa-S to about 6mPa-S at 22° C.

As used herein, the term “osmolality” refers to a measure of soluteconcentration, defined as the number of mmole of solute per kg ofsolution. A desired level of osmolality can be achieved by the additionof one or more stabilizer such as a sugar or sugar alcohol including,but not limited to, mannitol, dextrose, glucose, trehalose, and/orsucrose. Additional stabilizers that are suitable for providingosmolality are described in references such as the handbook ofPharmaceutical Excipients (Fourth Edition, Royal Pharmaceutical Societyof Great Britain, Science & Practice Publishers) or Remingtons: TheScience and Practice of Pharmacy (Nineteenth Edition, Mack PublishingCompany).

As used herein, the term “about” refers to +/−10% of the unit valueprovided. As used herein, the term “substantially” refers to thequalitative condition of exhibiting a total or approximate degree of acharacteristic or property of interest. One of ordinary skill in thebiological arts will understand that biological and chemical phenomenararely, if ever, achieve or avoid an absolute result. The termsubstantially is therefore used herein to capture the potential lack ofcompleteness inherent in many biological and chemical phenomena. As usedherein, the terms “isosmotic” and “isotonic” are used interchangeablywith the terms “substantially isosmotic,” and “substantially isotonic”and refer to formulations characterized by having an osmotic pressurethat is the same as or at least substantially equivalent to the osmoticpressure of another solution, which is achieved by formulations whereinthe total concentration of solutes, including both permeable andimpermeable solutes, in the formulation are the same as or at leastsubstantially equivalent to the total number of solutes in anothersolution. Thus, while it will be appreciated by those of skill in theart that “isosmotic” and “isotonic” formulations that are used for invivo administration generally have an osmolality ranging from about 270mmol/kg to about 310 mmol/kg, in the context of the low viscosityformulations of the present disclosure, the terms “isosmotic,”“isotonic,” “substantially isosmotic,” and “substantially isotonic” areused interchangeably to refer to formulations having an osmolalityranging from about 240 mmol/kg to about 380 mmol/kg, or from about 270mmol/kg to about 370 mmol/kg, or from about 300 mmol/kg to about 330mmol/kg.

The presently disclosed low viscosity, substantially isosmotic antibodyand other protein formulations contain from about 0 mM to about 30 mMhistidine; from about 50 ppm to about 200 ppm of a non-ionic surfactantsuch as, for example, polysorbate (Tween®) 80 or polysorbate (Tween®)20; from about 88 mM to about 292 mM of a sugar or sugar alcohol suchas, for example, mannitol, dextrose, glucose, trehalose, and/or sucrose;from about 0 mM to about 50 mM arginine; from about 0 mM to about 50 mMlysine; from about 0 mM to about 133 mM glycine or alanine; from about 0mM to about 10 mM methionine; and from about 1 mg/ml to about 150 mg/mlof a protein at a pH from about pH 4 to about pH 6. The formulationsdisclosed herein exhibit a viscosity ranging viscosity ranging fromabout 1 mPa-S to about 20 mPa-S at 22° C., or from about 1 mPa-S toabout 15 mPa-S at 22° C., or from about 1 mPa-S to about 10 mPa-S at 22°C. or from about 1 mPa-S to about 8 mPa-S at 22° C. or from about 1mPa-S to about 6 mPa-S at 22° C. and osmolality ranging from about 240to about 380 mmol/kg.

In these formulations, histidine—is a buffer agent, that can be used tomaintain the formulation pH from about pH 4 to about pH 6.0, or about pH5 to about pH 6, such as about pH 5, about pH 5.5, or about pH 6. Sugaror sugar alcohols, such as mannitol, dextrose, glucose, trehalose,and/or sucrose, are used separately or in combination, both ascryo-protectants and a stabilizer the antibody in liquid formulations aswell as during and after lyophilization. Non-ionic surfactants such aspolysorbates, including polysorbate 20 and polysorbate 80; polyoxamers,including poloxamer 184 and 188; Pluronic® polyols; and otherethylene/polypropylene block polymers, stabilize the antibody duringprocessing and storage by reducing interfacial interaction and preventprotein from adsorption. Arginine is a protein solubilizer and also astabilizer that reduces antibody and other protein aggregation, such asaTFPI mAb aggregation, and glycation. Methionine is an antioxidant thatprevents antibody oxidation during processing and storage.

Sugars and inorganic salts are commonly used as protein stabilizers;however, both sugars and inorganic salts are also effective tonicityagents. If a formulation requires a high concentration of one or moresugars to stabilize a protein, the inorganic salt concentration shouldbe zero or kept very low in order to maintain the formulation'sosmolality such that injection pain is reduced upon administration.Quite surprisingly, it was found that sodium chloride increased theturbidity of antibody formulations. Consequently, inorganic salts aresubstantially excluded from addition to the formulations describedherein. These non-salt formulations maintain the osmolality of theantibody and other protein formulations with increased stability, andreduced phase change, such as precipitation or aggregation.

As used herein, the term “salt” refers to inorganic salts, which includebut not limited to sodium chloride (NaCl), sodium sulfate (Na₂SO₄),sodium thiocyanate (NaSCN), magnesium chloride (MgCl), magnesium sulfate(MgSO₄), ammonium thiocyanate (NH₄SCN), ammonium sulfate ((NH₄)₂SO₄),ammonium chloride (NH₄Cl), calcium chloride (CaCl₂), calcium sulfate(CaSO₄), zinc chloride (ZnCl₂) and the like, or combinations thereof.The antibody and other protein formulations disclosed herein arecharacterized by a substantial absence of added salt and are, therefore,referred to herein as salt-free antibody and/or protein formulations. Itwill be understood by those of skill in the art that the presence ofinorganic salts within the presently disclosed formulations that areintroduced by pH adjustment are not considered to be added salts andsuch inorganic salts, if present in a formulation according to thepresent disclosure, should not exceed a concentration of about 2 mM.

As used herein, the term “surfactant” includes non-ionic surfactantsincluding, without limitation, polysorbates, such as polysorbate 20 or80, and the polyoxamers, such as poloxamer 184 or 188, Pluronic®polyols, and other ethylene/polypropylene block polymers. Amounts ofsurfactants effective to provide stable antibody and other proteinformulations are usually in the range of 50 ppm to 200 ppm. The use ofnon-ionic surfactants permits the formulations to be exposed to shearand surface stresses without causing denaturation of the antibody orother protein, and also reduce the adsorption on the surfaces duringprocessing and storage. The formulations disclosed herein include,without limitation, formulations having one or more non-ionicsurfactant(s) including, for example, one or more polysorbate(s), suchas polysorbate 20 or 80; one or more polyoxamers, such as poloxamer 184or 188; one or more Pluronic® polyol(s); and/or one or moreethylene/polypropylene block polymer(s). Exemplified herein areformulations having a polysorbate, such as polysorbate 20 (Tween® 20) orpolysorbate 80 (Tween® 80).

As used herein, the term “protein” refers to amino acid polymers thatcontain at least five constituent amino acids that are covalently joinedby peptide bonds. The constituent amino acids can be from the group ofamino acids that are encoded by the genetic code, which include:alanine, valine, leucine, isoleucine, methionine, phenylalanine,tyrosine, tryptophan, serine, threonine, asparagine, glutamine,cysteine, glycine, proline, arginine, histidine, lysine, aspartic acid,and glutamic acid. As used herein, the term “protein” is synonymous withthe related terms “peptide” and “polypeptide”.

As used herein, the term “antibody” refers to a class of proteins thatare generally known as immunoglobulins Antibodies include full-lengthmonoclonal antibodies (mAb), such as IgG₂ monoclonal antibodies, whichinclude immunoglobulin Fc regions. The term antibody also includesbispecific antibodies, diabodies, single-chain molecules, and antibodyfragments such as Fab, F(ab′)₂, and Fv.

As used herein, the term “anti-TFPI antibody” refers to an antibodyhaving binding specificity against the human TFPI protein as well asfragments and variants of the human TFPI protein. Anti-TFPI antibodiespresented herein can be IgG2 antibodies and include anti-TFPI IgG2monoclonal antibodies, such as chimeric, humanized, and fully-humananti-TFPI IgG2 monoclonal antibodies. Anti-TFPI antibodies areexemplified in the present disclosure by human anti-TFPI IgG2 monoclonalantibodies having a light chain comprising the sequence presented hereinas SEQ ID NO: 1 and/or a heavy chain presented herein as SEQ ID NO: 2.Other anti-TFPI monoclonal antibodies, including full-length antibodiesand antigen binding fragments and variants thereof, that are alsosuitable for use in the formulations disclosed herein are presented inPCT Patent Publication NOs. WO 2011/109452 and WO 2010/017196, both ofwhich are incorporated by reference herein in their entirety.

“Monoclonal antibodies” are characterized by having specificity for asingle antigenic determinant. Monoclonal antibodies can, for example, bemade by the hybridoma method described by Kohler and Milstein, Nature256:495 (1975) or by recombinant DNA methods such as those described inU.S. Pat. No. 4,816,567. Monoclonal antibodies can also be isolated fromphage display libraries using the techniques such as those described inClackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol.Biol. 222:581-597 (1991).

Monoclonal antibodies include “chimeric monoclonal antibodies” wherein aportion of a heavy and/or light chain includes sequences from antibodiesderived from one species, while the remainder of the antibody, includingthe Fc region, includes sequences from antibodies derived from a secondspecies, typically the second species is human. See, e.g., U.S. Pat. No.4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855(1984).

Monoclonal antibodies also include “humanized monoclonal antibodies”wherein one or more complementarity determining region (CDR) from aheavy and/or light chain sequence from antibodies derived from onespecies replace one or more CDR from a heavy and/or light chain sequencefrom antibodies derived from a second species, typically the secondspecies is human. The process of “humanization” is usually applied tomonoclonal antibodies developed for administration to humans. See, e.g.,Riechmann et al., Nature 332(6162):323-27 (1988) and Queen et al., Proc.Natl. Acad. Sci. USA 86(24):10029-33 (1989).

Monoclonal antibodies also include “fully-human monoclonal antibodies”wherein the entire heavy and light chain sequences are derived fromhuman antibody sequences. Fully-human monoclonal antibodies can begenerated by phage display technologies and can be isolated from micethat have been genetically engineered to express the human antibodyrepertoire. See, e.g., McCafferty et al., Nature 348(6301):552-554(1990), Marks et al., J. Mol. Biol. 222(3):581-597 (1991), and Carmenand Jermutus, Brief Funct. Genomic Proteomic 1(2):189-203 (2002).

As used herein, the term “Pharmaceutically effective amount” of anantibody or other protein formulation refers to an amount of theformulation that provides therapeutic effect in an administrationregimen. The antibody and protein formulations disclosed hereintypically include an antibody or other protein at a concentrationranging from about 1 mg/ml to about 150 mg/ml, or from about 1 mg/ml toabout 100 mg/ml, or from about 1 mg/ml to about 50 mg/ml, or from about1 mg/ml to about 20 mg/ml, or from about 1 mg/ml to about 10 mg/ml, orfrom about 10 mg/ml to about 20 mg/ml, or from about 20 mg/ml to about150 mg/ml, or from about 50 mg/ml to about 150 mg/ml, or from about 60mg/ml to about 150 mg/ml, or from about 70 mg/ml to about 150 mg/ml, orfrom about 80 mg/ml to about 150 mg/ml, or from about 90 mg/ml to about150 mg/ml, or from about 100 mg/ml to about 150 mg/ml, or from about 120mg/ml to about 150 mg/ml, or from about 140 mg/ml to about 150 mg/ml,Within some aspects the concentration of protein or antibody withinthese formulations is about 150 mg/ml. When administered subcutaneously,such formulations are typically administered in a volume of less thanabout 2.0 ml, or about 1.5 ml, or about 1 ml, or about 0.5 ml perinjection site.

Within certain aspects, the antibody or other protein formulationcontains about 30 mM histidine, about 100 ppm Tween 80, about 292 mMsucrose, about 20 mg/ml antibody or other protein at a pH ranging fromabout pH 5.0 to about pH 6.0. Within related aspects, the antibody andother protein formulation also contains from about 30 mM to about 50 mMarginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 234 mM sucrose,about 50 mg/ml antibody or other protein at a pH ranging from about pH5.0 to about pH 6.0. Within related aspects, the antibody and otherprotein formulation also contains from about 30 mM to about 50 mMarginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 234 mM sucrose,about 100 mg/ml antibody or other protein at a pH ranging from about pH5.0 to about pH 6.0. Within related aspects, the antibody and otherprotein formulation also contains from about 30 mM to about 50 mMarginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 88 mM sucrose, about133 mM glycine, about 100 mg/ml antibody or other protein at a pHranging from about pH 5.0 to about pH 6.0. Within related aspects, theantibody and other protein formulation also contains from about 30 mM toabout 50 mM arginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 20, about 88 mM sucrose, about133 mM glycine, about 100 mg/ml antibody or other protein at a pHranging from about pH 5.0 to about pH 6.0. Within related aspects, theantibody and other protein formulation also contains from about 30 mM toabout 50 mM arginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 200 ppm Tween 20, about 88 mM sucrose,about 133 mM glycine, about 100 mg/ml antibody or other protein at a pHranging from about pH 5.0 to about pH 6.0. Within related aspects, theantibody and other protein formulation also contains from about 30 mM toabout 50 mM arginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 88 mM sucrose, about100 mg/ml antibody or other protein at a pH ranging from about pH 5.0 toabout pH 6.0. Within related aspects, the antibody and other proteinformulation also contains from about 10 mM to about 50 mM arginine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 88 mM sucrose, about133 mM glycine, about 10 mM arginine, about 100 mg/ml antibody or otherprotein at a pH ranging from about pH 5.0 to about pH 6.0. Withinrelated aspects, the antibody and other protein formulation alsocontains from about 0 mM to about 10 mM methionine.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm Tween 80, about 88 mM sucrose, about133 mM glycine, about 30 mM lysine, about 100 mg/ml antibody or otherprotein at a pH ranging from about pH 5.0 to about pH 6.0.

Within other aspects, the antibody or other protein formulation containsabout 10 mM histidine, about 75 ppm about Tween 80, about 234 mMsucrose, about 30 mM arginine, about 100 mg/ml antibody or other proteinat a pH ranging from about pH 5.0 to about pH 6.0. Within relatedaspects, the antibody and other protein formulation also contains fromabout 0 mM to 10 mM methionine.

Exemplified herein are antibody formulations wherein the antibodiesinclude IgG2 antibodies, such as anti-tissue factor pathway inhibitorantibodies (aTFPI Abs), including the human IgG2 monoclonal antibodyhaving a light chain comprising the sequence of SEQ ID NO: 1 and a heavychain comprising the sequence of SEQ ID NO: 2.

Thus, the present disclosure provides anti-TFPI mAb formulations,including anti-TFPI IgG₂ mAb formulations, wherein the anit-TFPI mAb issoluble at high protein concentrations. Typically, the anti-TFPI mAb inthe formulations disclosed herein remain soluble at concentrations ofbetween about 1 mg/ml and about 150 mg/ml and remain stable underisosmotic storage conditions and exhibit reduced viscosity as comparedto currently available antibody formulations.

The anti-TFPI antibody having a light chain comprising the sequence ofSEQ ID NO: 1 and a heavy chain comprising the sequence of SEQ ID NO: 2is an IgG₂ antibody that blocks tissue factor pathway inhibitor (TFPI).Since TFPI down-regulates extrinsic coagulation, anti-TFPI antibodiescan promote extrinsic pathway-driven coagulation by blocking TFPI,thereby bypassing FVIII or FIX deficiencies in the intrinsic pathway forhemophilia treatment. The salt free anti-TFPI antibody formulationspresented herein can be administrated to the patients via subcutaneousinjection or other injection routes.

As part of the present disclosure, it was found that solubility andstability of anti-TFPI antibodies was affected by excipients. Thesolubility of anti-TFPI antibody increases with the decrease of NaClconcentrations. In the absence of NaCl, the solubility of anti-TFPIantibody is higher than formulations that include NaCl. In addition, itwas found that positively changed amino acids, such as arginine andlysine, could improve the stability anti-TFPI antibody and that pHgreatly affected the anti-TFPI antibody solubility. The turbidity of theantibody solution increased with increases in pH; however, theprecipitation was reversible when pH was decreased. The optimal pH forstabilizing the anti-TFPI antibodies presented herein ranges from aboutpH 4 to about pH 6 or from about pH 5 to about pH 6, such as about pH 5,about pH 5.5, or about pH 6.

Exemplified herein are formulations, as recited above, wherein theantibody is an anti-TFPI antibody (aTFPI Ab). In at least one aspect,the anti-TFPI antibody is a human IgG2 monoclonal antibody. For example,the human anti-TFP1 IgG2 monoclonal antibody includes the antibody thatcontains a light chain having the amino acid sequence presented in SEQID NO: 1 and a heavy chain having the amino acid sequence presented inSEQ ID NO: 2.

Heavy and Light Chain Sequences of an Exemplary Human Anti-TFPI IgG2Monoclonal Antibody Sequence Amino Acid Sequence Identifier (NH₃-COOH)SEQ ID NO: 1 SYELTQPPSV SVSPGQTARI TCSGDNLPKYYAHWYQQKPG QAPVVVIFYD VNRPSGIPER FSGSNSGNTA TLTISGTQAM DEADYYCQAWWSSTPVFGGG TKLTVLGQPK AAPSVTLFPP SSEELQANKA TLVCLISDFY PGAVTVAWKADSSPVKAGVE TTTPSKQSNN KYAASSYLSL TPEQWKSHRS YSCQVTHEGS TVEKTVAPTE CSSEQ ID NO: 2 EVQLVESGGG LVQPGGSLRL SCAASGFTFSSYGMDWVRQA PGKGLEWVSS IRGSRGSTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAEDTAVYYCARLY RYWFDYWGQG TLVTVSSAST KGPSVFPLAP CSRSTSESTA ALGCLVKDYFPEPVTVSWNS GALTSGVHTF PAVLQSSGLY SLSSVVTVPS SNFGTQTYTC NVDHKPSNTKVDKTVERKCC VECPPCPAPP VAGPSVFLFP PKPKDTLMIS RTPEVTCVVV DVSHEDPEVQFNWYVDGVEV HNAKTKPREE QFNSTFRVVS VLTVVHQDWL NGKEYKCKVS NKGLPAPIEKTISKTKGQPR EPQVYTLPPS REEMTKNQVS LTCLVKGFYP SDIAVEWESN GQPENNYKTTPPMLDSDGSF FLYSKLTVDK SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PG

The present disclosure also provides methods for the treatment of adisorder in a patient, comprising the administration to the patient of atherapeutically effective amount of one or more formulations describedherein. For example, provided are methods for the treatment of adisorder in a patient, comprising the administration to the patient of atherapeutically effective amount of an antibody or other proteinformulation that contains from about 0 mM to about 30 mM histidine and,from about 50 ppm to about 200 ppm polysorbate (Tween®) 80 orpolysorbate (Tween®) 20, from about 88 mM to about 292 mM sucrose, fromabout 0 mM to about 50 mM arginine, from about 0 mM to about 50 mMlysine, from about 0 mM to about 133 mM glycine or alanine, from about 0mM to about 10 mM methionine, and from about 1 mg/ml to about 150 mg/mlof a protein at a pH ranging from about pH 4.0 to about pH 6.0. Withinat least one aspect of these methods, the antibody or other proteinformulation can be administered intravenously. Within other aspects ofthese methods, the antibody or other protein formulation can beadministered subcutaneously. Within other aspects of these methods, theantibody or other protein formulation can be administeredintramuscularly.

Within related aspects, the present disclosure provides methods for thetreatment of hemophilia A or hemophilia B in a patient, comprising theadministration to the patient of a therapeutically effective amount ofan anti-TFPI antibody formulation that contains from about 0 mM to about30 mM histidine and, from about 50 ppm to about 200 ppm polysorbate(Tween®) 80 or polysorbate (Tween®) 20, from about 88 mM to about 292 mMsucrose, from about 0 mM to about 50 mM arginine, from about 0 mM toabout 50 mM lysine, from about 0% (0 mM) to about 1% (133 mM) glycine,from about 0 mM to about 10 mM methionine, and from about 1 mg/ml toabout 150 mg/ml of a protein at a pH ranging from about pH 4.0 to aboutpH 6.0. Within at least one aspect of these methods, the anti-TFPIantibody formulation can be administered intravenously. Within otheraspects of these methods, the anti-TFPI antibody formulation can beadministered subcutaneously. Within other aspects of these methods, theantibody or other protein formulation can be administeredintramuscularly.

According to certain aspects of these methods for the treatment ofhemophilia A or hemophilia B in a patient, the anti-TFPI antibody is ahuman anti-TFP1 IgG2 monoclonal antibody such as, for example, the humananti-TFPI IgG2 monoclonal antibody that contains a light chain havingthe amino acid sequence presented in SEQ ID NO: 1 and a heavy chainhaving the amino acid sequence presented in SEQ ID NO: 2.

For the purpose of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural and vice versa. In the event thatany definition set forth below conflicts with the usage of that word inany other document, including any document incorporated herein byreference, the definition set forth below shall always control forpurposes of interpreting this specification and its associated claimsunless a contrary meaning is clearly intended (for example in thedocument where the term is originally used). The use of “or” means“and/or” unless stated otherwise. The use of “a” herein means “one ormore” unless stated otherwise or where the use of “one or more” isclearly inappropriate. The use of “comprise,” “comprises,” “comprising,”“include,” “includes,” and “including” are interchangeable and notintended to be limiting. Furthermore, where the description of one ormore embodiments uses the term “comprising,” those skilled in the artwould understand that, in some specific instances, the embodiment orembodiments can be alternatively described using the language“consisting essentially of” and/or “consisting of.”

Aspects of the present disclosure may be further understood in light ofthe following examples, which should not be construed as limiting thescope of the present teachings in any way.

EXAMPLES Example 1 Effect of NaCl Concentration and pH on the Turbidityof Antibody Solutions

This Example discloses the effect of salt (NaCl) concentration and pH onthe turbidity of solutions containing an anti-TFPI human monoclonalantibody having a light chain with the amino acid sequence presented inSEQ ID NO: 1 and a heavy chain with the amino acid sequence presented inSEQ ID NO: 2.

The turbidity of solutions was measured by Nephelometry to quicklyevaluate the effects of salt concentrations and pH on aTFPI Absolutions. The formulation of anti-TFPI antibody used in this Examplecontained 10 mM acetate buffer, 88 mM sucrose, and 200 ppm Tween 80.NaCl concentrations were varied from 0 mM to 300 mM. The results ofNaCl-dependent turbidity measurements for anti-TFPI formulations at pH5.5 are presented in FIG. 1. These data demonstrated that the turbidityof the anti-TFPI mAb formulations increased significantly withincreasing concentration of NaCl. Increasing salt concentration from 0to 300 mM resulted in an increase of 72 FNU in turbidity value byNephelometry, which could be attributed to precipitation, aggregation orinsolubilization of aTFPI Ab in solution. As the result of this finding,solutions without sodium chloride were recommended for the presentlydisclosed anti-TFPI antibody formulations.

Without being bound by theory, it is believed that the increasedturbidity of the anti-TFPI mAb formulations with increasing NaClconcentration resulted from the neutralization of positive charges onthe anti-TFPI mAb arginine side-chains. The phase behavior of aTFPI mAbat different pH with the impact of monovalent salt (NaCl) explains whythe stable, soluble, non-salt, and substantial isosmolality aTFPI mAbformulations could be achieved.

The presently disclosed aTFPI mAb molecule has 116 amino acids (42arginines and 74 lysines) having side-chain carrying positive charges atthe pH below PI. This anti-TFPI antibody has a PI at ˜7.9. At a pH belowthe PI, such as pH 4-6, this anti-TFPI antibody has net positivecharges. The repulsion of the positive charges on this anti-TFPIantibody surface likely prevents protein-protein association betweenindividual molecules and, thereby, significantly increases solubility.It is hypothesized that the anion (Cl⁻) of salt binds to the guanidiniumgroup on arginine side-chains on the anti-TFPI antibody surface toneutralize the positive charges, which enhances protein-proteininteractions and, hence, causes lower solubility and solution turbidity.By shifting the pH to 4-6, the non-salt formulations that are describedherein were developed to achieve increased antibody solubility andstability (see, FIG. 1). In absence of salt, the concentration of otherstabilizers, such as sucrose, can be increased to >150 mM and <300 mMwithout compromising osmolality.

The effect of pH on the turbidity of anti-TFPI antibodies was alsostudied. As shown in FIG. 2, pH can also greatly affect the turbidity ofaTFPI Ab solution. When pH was increased from 4 to 6.5, the turbidity ofaTFPI Ab solution increased by 81 FNU. Further increasing pH to 7, theturbidity of solution was out of the range for accurate measurement.However, the formed precipitates in the solution were reversible when pHwas decreased. This result could be attributed to the surface chargeneutralization when pH was increased to the value close to the PI ofaTFPI Ab, since the PI value of aTFPI Ab is approximately 7.9. Accordingto this study, lower pH was preferred for aTFPI Ab formulations.However, low pH could cause tissue irritation during injection.Therefore, a neutral pH was preferred from patient compliance point ofview. Balancing these two factors, the optimum pH of aTFPI Abformulations is between pH 5 and pH 6.

Example 2 Anti-TFPI Antibody Formulations

Based upon the unexpected findings presented in Example 1, substantiallyisosmotic anti-TFPI Ab formulations were prepared without NaCl. Theseformulations typically employed high sucrose concentrations to helpstabilize the anti-TFPI Ab.

Frozen anti-TFPI antibody was thawed and reformulated by dialysisaccording to formulations presented in Table 1. The formulations wereprepared and were filtered with a 0.22 μm filter and filled in glasstubing vials and stoppered with rubber stoppers.

It was also found that in the absence of NaCl, and in the presence ofsucrose 88 mM to 292 mM and polysorbate 80 or polysorbate 20 (50-200ppm), the positive charged amino acids, such as arginine (10-50 mM), caneffectively inhibit aTFPI Ab from glycation.

TABLE 1 Anti-TFPI Antibody Formulations Formulation DesignationFormulation Composition PH5.0  20 mg/ml aTFPI mAb  30 mM histidine 292mM sucrose 100 ppm Tween 80 pH 5.0 PH5.5  20 mg/ml aTFPI mAb  30 mMhistidine 292 mM sucrose 100 ppm Tween 80 pH 5.5 PH6.0  20 mg/ml aTFPImAb  30 mM histidine 292 mM sucrose 100 ppm Tween 80 pH 6.0 6ARG  20mg/ml aTFPI mAb  30 mM histidine 292 mM sucrose 100 ppm Tween 80  50 mMarginine pH 6.0 PH5.5LC  50 mg/ml aTFPI Ab  10 mM histidine 234 mMsucrose  75 ppm Tween 80 pH 5.5 PH5.5LCARG  50 mg/ml aTFPI Ab  10 mMhistidine 234 mM sucrose  75 ppm Tween 80  30 mM arginine pH 5.5PH5.5HCARG 100 mg/ml aTFPI Ab  10 mM histidine 234 mM sucrose  75 ppmTween 80  30 mM arginine pH 5.5 PH6LCARG  50 mg/ml aTFPI Ab  10 mMhistidine 234 mM sucrose  75 ppm Tween 80  50 mM arginine pH at 6PH6LCARG_L 100 mg/ml aTFPI Ab  10 mM histidine 234 mM sucrose  75 ppmTween 80  30 mM arginine pH 6 3% STWN80 100 mg/ml aTFPI Ab  10 mMhistidine  88 mM sucrose  75 ppm Tween 80  30 mM arginine 133 mM glycinepH 5.5 3% STWN20_L 100 mg/ml aTFPI Ab  10 mM histidine  88 mM sucrose 75 ppm Tween 20  30 mM arginine 133 mM glycine pH 5.5 3% STWN20_H 100mg/ml aTFPI Ab  10 mM histidine  88 mM sucrose 200 ppm Tween 20  30 mMarginine 133 mM glycine pH at 5.5 5.5ARG10 100 mg/ml aTFPI Ab  10 mMhistidine  88 mM sucrose  75 ppm Tween 80  10 mM arginine 133 mM glycinepH 5.5 5.5ARG10MET 100 mg/ml aTFPI Ab  10 mM histidine  88 mM sucrose 75 ppm Tween 80  10 mM arginine  10 mM methionine 133 mM glycine pH 5.55.5LYS30 100 mg/ml aTFPI Ab  10 mM histidine  88 mM sucrose  75 ppmTween 80  30 mM lysine 133 mM glycine pH 5.5 PH5.5HCARGMET 100 mg/mlaTFPI Ab  10 mM histidine 234 mM sucrose  75 ppm Tween 80  30 mMarginine  10 mM methionine pH 5.5

Each of these anti-TFPI mAb formulations was analyzed by HPLC-SEC forprotein aggregation and degradation, LC-MS for aTFPI structural changes(glycation and oxidation), and nephlometry for turbidity assessment,viscometer for viscosity measurement, and osmolality instrument forosmolality measurement. The results for these analyses are presented inTable 2.

TABLE 2 Osmolality and Viscosity of Anti-TFPI Antibody FormulationsFormulation Viscosity at 22° C. Osmolality Composition (mPa-S) (mmol/kg)aTFPI 50 mg/ml 1.8 272 Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH5.5 aTFPI 50 mg/ml 1.6 339 Histidine 10 mM Sucrose 234 mM Tween 80 75ppm pH 5.5 Arginine 30 mM aTFPI 100 mg/ml 2.9 335 Histidine 10 mMSucrose 234 mM Tween 80 75 ppm pH 5.5 Arginine 30 mM aTFPI 50 mg/ml 2.3-373 Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine 50 mMaTFPI 100 mg/ml 4.6  353* Histidine 10 mM Sucrose 234 mM Tween 80 75 ppmpH 6.0 Arginine 30 mM aTFPI 100 mg/ml 4.1  282* Histidine 10 mM Sucrose88 mM Tween 80 75 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100mg/ml 4.5 282 Histidine 10 mM Sucrose 88 mM Tween 20 75 ppm pH 5.5Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml 4.5 278 Histidine 10 mMSucrose 88 mM Tween 20 200 ppm pH 5.5 Arginine 30 mM Glycine 133 mMaTFPI 100 mg/ml —  263* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH5.5 Arginine 10 mM Glycine 133 mM aTFPI 100 mg/ml —  268* Histidine 10mM Sucrose 88 mM Tween 80 75 ppm pH 5.5 Arginine 10 mM Methionine 10 mMGlycine 133 mM aTFPI 100 mg/ml —  288* Histidine 10 mM Sucrose 88 mMTween 80 75 ppm pH 5.5 Lysine 30 mM Glycine 133 mM aTFPI 100 mg/ml 3.0341 Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 5.5 Arginine 30 mMMethionine 10 mM *Estimated osmolality based on calculations

Example 3 The Effect of Arginine on Anti-TFPI Ab Glycation

The Example demonstrates that anti-TFPI Ab formulations containingarginine exhibit reduced antibody glycation as compared to anti-TFPI Abformulations without added arginine.

Anti-TFPI Ab formulations, with and without arginine, at pH 6 werestored at 40° C. for 14 days and tested by LC-MS. The results presentedin Table 3 demonstrate that the positively-charged amino acid argininereduces glycation of anti-TFPI Ab, possibly due to the unique structureof aTFPI Ab, to the level found in a reference standard.

TABLE 3 Arginine Reduces Anti-TFPI mAb Glycation Formulation FormulationDesignation Composition LC-MS Profile PH6.0  20 mg/ml aTFPI mAbGlycation  30 mM Histidine 292 mM Sucrose 100 ppm Tween 80 pH 6.0 6ARG 20 mg/ml aTFPI mAb Comparable to  30 mM Histidine Reference Standard292 mM Sucrose 100 ppm Tween 80  50 mM Arginine pH 6.0

It can be concluded based on the results of this study that thestability of aTFPI Ab is highly impacted by formulation pH and theoptimum pH for the stability of the formulation is between pH 5 and pH 6when IV, IM and subcutaneous injection are considered. Arginine appearedto be able to prevent aTFPI Ab glycation. The formulation developmentand stability studies presented in Example 4 and Example 5 were designedbased on these findings.

Example 4 Stability of Anti-TFPI Ab Formulations

The HPLC-SEC results and the LC-MS results of anti-TFPI Ab formulationsare summarized in Tables 4-7.

TABLE 4 Stability of Anti-TFPI Ab Lyophilization Formulations after 3Months at 5° C. HPLC-SEC Average rate of Formulation aggregationformation LC-MS Composition (%/day)² (3 month) aTFPI 50 mg/ml 0 YHistidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 5.5 aTFPI 50 mg/ml 0 YHistidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 5.5 Arginine 30 mMaTFPI 100 mg/ml 0 Y Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH5.5 Arginine 30 mM aTFPI 50 mg/ml 0 Y Histidine 10 mM Sucrose 234 mMTween 80 75 ppm pH 6.0 Arginine 50 mM aTFPI 100 mg/ml 0 Y* Histidine 10mM Sucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine 30 mM aTFPI 100 mg/ml0 Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH 5.5 Arginine 30 mMGlycine 133 mM aTFPI 100 mg/ml  0¹ Y¹ Histidine 10 mM Sucrose 88 mMTween 20 75 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml  0¹Y¹ Histidine 10 mM Sucrose 88 mM Tween 20 200 ppm pH 5.5 Arginine 30 mMGlycine 133 mM ¹The calculation was based on 2 month value. ²If the rateis negative, zero was entered. Y: Comparable to the reference standardY*: Comparable to the reference standard, with minor modification

TABLE 5 Stability of Anti-TFPI Ab Lyophilization Formulations after 3Months at 40° C. HPLC-SEC Average rate of Formulation aggregationformation LC-MS Composition (%/day)² (3 month) aTFPI 50 mg/ml 0.01Glycated Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 5.5 aTFPI 50mg/ml 0 Y Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 5.5 Arginine30 mM aTFPI 100 mg/ml 0.01 Y Histidine 10 mM Sucrose 234 mM Tween 80 75ppm pH 5.5 Arginine 30 mM aTFPI 50 mg/ml 0 Y Histidine 10 mM Sucrose 234mM Tween 80 75 ppm pH 6.0 Arginine 50 mM aTFPI 100 mg/ml 0.01 Y*Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine 30 mMaTFPI 100 mg/ml 0.01 Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH5.5 Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml 0.04¹ Y¹ Histidine 10mM Sucrose 88 mM Tween 20 75 ppm pH 5.5 Arginine 30 mM Glycine 133 mMaTFPI 100 mg/ml 0.04¹ Y¹ Histidine 10 mM Sucrose 88 mM Tween 20 200 ppmpH 5.5 Arginine 30 mM Glycine 133 mM ¹The calculation was based on 2month value. ²If the rate is negative, zero was entered. Y: Comparableto the reference standard Y*: Comparable to the reference standard, withminor modification

TABLE 6 Liquid Formulations at 5° C. for 3 Months HPLC-SEC Average rateof aggregation LC-MS Formulation Composition formation (%/day)² (3month) aTFPI 50 mg/ml 0 Y Histidine 10 mM Sucrose 234 mM Tween 80 75 ppmpH 5.5 aTFPI 50 mg/ml 0 Y Histidine 10 mM Sucrose 234 mM Tween 80 75 ppmpH 5.5 Arginine 30 mM aTFPI 100 mg/ml 0 Y Histidine 10 mM Sucrose 234 mMTween 80 75 ppm pH 5.5 Arginine 30 mM aTFPI 50 mg/ml 0 Y Histidine 10 mMSucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine 50 mM aTFPI 100 mg/ml  0.01 Y* Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine30 mM aTFPI 100 mg/ml   0.01 Y* Histidine 10 mM Sucrose 88 mM Tween 8075 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml  0¹ Y¹Histidine 10 mM Sucrose 88 mM Tween 20 75 ppm pH 5.5 Arginine 30 mMGlycine 133 mM aTFPI 100 mg/ml  0¹ Y¹ Histidine 10 mM Sucrose 88 mMTween 20 200 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml 0Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH 5.5 Arginine 10 mMaTFPI 100 mg/ml 0 Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH5.5 Arginine 10 mM Methionine 10 mM aTFPI 100 mg/ml   0.01 Y* Histidine10 mM Sucrose 88 mM Tween 80 75 ppm pH 5.5 Lysine 30 mM ¹The calculationwas based on 2 months value. ²If the rate is negative, zero was entered.Y: Comparable to the reference standard Y*: Comparable to the referencestandard, with minor modification

TABLE 7 Liquid Formulations at 40° C. for 3 Months HPLC-SEC Average rateof Formulation aggregation formation LC-MS Composition (%/day) (3 month)aTFPI 50 mg/ml 0.17 Glycated Histidine 10 mM Sucrose 234 mM Tween 80 75ppm pH 5.5 aTFPI 50 mg/ml 0.02 Y* Histidine 10 mM Sucrose 234 mM Tween80 75 ppm pH 5.5 Arginine 30 mM aTFPI 100 mg/ml 0.04 Y* Histidine 10 mMSucrose 234 mM Tween 80 75 ppm pH 5.5 Arginine 30 mM aTFPI 50 mg/ml 0.03Y* Histidine 10 mM Sucrose 234 mM Tween 80 75 ppm pH 6.0 Arginine 50 mMaTFPI 100 mg/ml 0.04 Y* Histidine 10 mM Sucrose 234 mM Tween 80 75 ppmpH 6.0 Arginine 30 mM aTFPI 100 mg/ml 0.04 Y* Histidine 10 mM Sucrose 88mM Tween 80 75 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100 mg/ml0.04¹ Y*¹ Histidine 10 mM Sucrose 88 mM Tween 20 75 ppm pH 5.5 Arginine30 mM Glycine 133 mM aTFPI 100 mg/ml 0.040¹ Y*¹ Histidine 10 mM Sucrose88 mM Tween 20 200 ppm pH 5.5 Arginine 30 mM Glycine 133 mM aTFPI 100mg/ml 0.07 Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH 5.5Arginine 10 mM aTFPI 100 mg/ml 0.06 Y* Histidine 10 mM Sucrose 88 mMTween 80 75 ppm pH 5.5 Arginine 10 mM Methionine 10 mM aTFPI 100 mg/ml0.06 Y* Histidine 10 mM Sucrose 88 mM Tween 80 75 ppm pH 5.5 Lysine 30mM ¹The calculation was based on 2 month value. Y: Comparable to thereference standard Y*: Comparable to the reference standard, with minormodification

What is claimed is:
 1. A salt-free protein or antibody formulation,comprising: a. 10 mM to 30 mM histidine; b. 50 ppm to 200 ppm of anon-ionic surfactant; c. 88 mM to 292 mM of a sugar or sugar alcoholselected from mannitol, dextrose, glucose, trehalose, and sucrose; d. 0mM to 50 mM arginine; e. 0 mM to 50 mM lysine; f. 0 mM to 133 mM glycineor alanine; g. 0 mM to 10 mM methionine; and h. 1 mg/ml to 150 mg/ml ofa protein or an antibody; wherein said protein or antibody formulationhas a pH ranging from pH 4.0 to pH 6.0 and wherein said proteinformulation contains substantially no inorganic salt.
 2. The salt-freeprotein or antibody formulation of claim 1 wherein said formulation hasa viscosity ranging from 1 to 8 mPa-S at 22° C.
 3. The salt-free proteinor antibody formulation of claim 1 wherein said formulation has andosmolality ranging from 240 to 380 mmol/kg.
 4. The salt-free protein orantibody formulation of any of claim 1 wherein said non-ionic surfactantis a polysorbate selected from polysorbate 20 and polysorbate
 80. 5. Thesalt-free protein or antibody formulation of claim 1 wherein said sugaris sucrose.
 6. The salt-free protein or antibody formulation of claim 1comprising between 10 mM and 50 mM arginine.
 7. The salt-free protein orantibody formulation of claim 1, comprising: a. 30 mM histidine, b. 100ppm polysorbate 80, c. 292 mM sucrose, and d. 20 mg/ml protein orantibody; wherein said protein or antibody formulation has a pH rangingfrom pH 5.0 to pH 6.0.
 8. The salt-free protein or antibody formulationof claim 1, comprising: a. 10 mM histidine, b. 75 ppm polysorbate 80, c.234 mM sucrose, d. 50 mg/ml protein or antibody; wherein said protein orantibody formulation has a pH ranging from pH 5.0 to pH 6.0.
 9. Thesalt-free protein or antibody formulation of claim 1, comprising: a. 10mM histidine, b. 75 ppm polysorbate 80, c. 234 mM sucrose, and d. 100mg/ml protein or antibody; wherein said protein or antibody formulationhas a pH ranging from pH 5.0 to pH 6.0.
 10. The salt-free protein orantibody formulation of claim 1, comprising: a. 10 mM histidine, b. 75ppm polysorbate 80, c. 88 mM sucrose, d. 133 mM glycine, and e. 100mg/ml protein or antibody; wherein said protein or antibody formulationhas a pH ranging from pH 5.0 to pH 6.0.
 11. The salt-free protein orantibody formulation of claim 1, comprising: a. 10 mM histidine, b. 75ppm polysorbate 20, c. 88 mM sucrose, d. 133 mM glycine, and e. 100mg/ml protein or antibody; wherein said protein or antibody formulationhas a pH ranging from pH 5.0 to pH 6.0.
 12. The salt-free protein orantibody formulation of claim 1, comprising: a. 10 mM histidine, b. 200ppm polysorbate 20, c. 88 mM sucrose, d. 133 mM glycine, and e. 100mg/ml protein or antibody; wherein said protein or antibody formulationhas a pH ranging from pH 5.0 to pH 6.0.
 13. The salt-free protein orantibody formulation of claim 1, comprising: a. 10 mM histidine, b. 75ppm polysorbate 80, c. 88 mM sucrose, and d. 100 mg/ml protein orantibody; wherein said protein or antibody formulation has a pH rangingfrom pH 5.0 to pH 6.0.
 14. The salt-free protein or antibody formulationof claim 1, comprising: a. 10 mM histidine, b. 75 ppm polysorbate 80, c.88 mM sucrose, d. 10 mM arginine, and e. 100 mg/ml protein or antibody;wherein said protein or antibody formulation has a pH ranging from pH5.0 to pH 6.0.
 15. The salt-free protein or antibody formulation ofclaim 1, comprising 5 mM to 10 mM methionine.
 16. The salt-free proteinor antibody formulation of claim 1, comprising: a. 10 mM histidine, b.75 ppm polysorbate 80, c. 88 mM sucrose, d. 30 mM lysine, and e. 100mg/ml protein or antibody; wherein said protein or antibody formulationhas a pH ranging from pH 5.0 to pH 6.0.
 17. A salt-free anti-TFPIantibody formulation, comprising: a. 10 mM to 30 mM histidine, b. 50 ppmto 200 ppm of a non-ionic surfactant, c. 88 mM to 292 mM of a sugar orsugar alcohol selected from mannitol, dextrose, glucose, trehalose andsucrose, d. 0 mM to 50 mM arginine, e. 0 mM to 50 mM lysine, f. 0 mM to133 mM glycine or alanine g. 0 mM to 10 mM methionine, and h. 1 mg/ml to150 mg/ml of anti-TFPI antibody; wherein said anti-TFPI antibodyformulation has a pH of pH 4.0 to pH 6.0 and wherein said anti-TFPIantibody formulation contains substantially no inorganic salt.
 18. Thesalt-free anti-TFPI antibody formulation of claim 17 wherein saidformulation has a viscosity ranging from 1 to 8 mPa-S at 22° C.
 19. Thesalt-free anti-TFPI antibody formulation of claim 17 wherein saidformulation has and osmolality ranging from 240 to 380 mmol/kg.
 20. Thesalt-free anti-TFPI antibody formulation of claims 17 wherein saidnon-ionic surfactant is a polysorbate selected from polysorbate 20 andpolysorbate
 80. 21. The salt-free anti-TFPI antibody formulation ofclaim 17 wherein said sugar is sucrose.
 22. The salt-free anti-TFPIantibody formulation of claim 17, comprising: a. 10 mM histidine, b. 75ppm Tween 80, c. 234 mM sucrose, d. 30 mM arginine, and e. 10 mMmethionine, and f. 100 mg/ml anti-TFPI antibody; wherein said anti-TFPIantibody formulation has a pH of 5.5.
 23. The salt-free anti-TFPIantibody formulation of claim 17 wherein said anti-TFPI antibody is ahuman IgG₂ monoclonal antibody.
 24. The salt-free anti-TFPI antibodyformulation of claim 17 wherein said human IgG₂ monoclonal antibodycomprises a light chain comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 2.25. A method for the treatment of a disorder in a patient, said methodcomprising administering to said patient a therapeutically effectiveamount of a protein or antibody formulation comprising between 10 mM and30 mM histidine, between 50 ppm and 200 ppm polysorbate 80 orpolysorbate 20, between 88 mM and 292 mM sucrose, between 0 mM and 50 mMarginine, between 0 mM and 50 mM lysine, between 0 mM and 133 mMglycine, between 0 mM and 10 mM methionine, and between 1 mg/ml and 150mg/ml of a protein or antibody at a pH of between pH 4.0 and pH 6.0,wherein said protein or antibody formulation contains substantially noinorganic salt.
 26. The method of claim 25 wherein said protein orantibody formulation is administered intravenously, subcutaneously, orintramuscularly.
 27. A method for the treatment of hemophilia A orhemophilia B in a patient, said method comprising administering to saidpatient a therapeutically effective amount of an anti-TFPI antibodyformulation that contains between 10 mM and 30 mM histidine, between 50ppm and 200 ppm polysorbate 80 or polysorbate 20, between 88 mM and 292mM sucrose, between 0 mM and 50 mM arginine, between 0 mM and 50 mMlysine, between 0 mM and 133 mM glycine, between 0 mM and 10 mMmethionine, and between 1 mg/ml and 150 mg/ml of a protein at a pH ofbetween pH 4.0 and pH 6.0, and wherein said anti-TFPI formulationcontains substantially no inorganic salt.
 28. The method of claim 27wherein said anti-TFPI antibody is a human IgG₂ monoclonal antibody. 29.The method of claim 28 wherein said human IgG₂ monoclonal antibodycomprises a light chain comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 2.30. The method of claim 44 wherein said anti-TFPI antibody formulationis administered intravenously, intramuscularly, or subcutaneously.